ArticleProtein science : a publication of the Protein Society2007
Structure-based design of robust glucose biosensors using a Thermotoga maritima periplasmic glucose-binding protein.
Article in Protein science : a publication of the Protein Society, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed.
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- Development of a Real-Time Pectic Oligosaccharide-Detecting Biosensor Using the Rapid and Flexible Computational Identification of Non-Disruptive Conjugation Sites (CINC) Biosensor Design Platform.Sensors (Basel, Switzerland) · 2022Article
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- Genome-Wide Analysis of Sugar Transporters Identifies theMicroorganisms · 2020Article
- A FRET-based biosensor for the quantification of glucose in culture supernatants of mL scale microbial cultivations.Microbial cell factories · 2019Article
- Periplasmic Binding Proteins in Thermophiles: Characterization and Potential Application of an Arginine-Binding Protein from Thermotoga maritima: A Brief Thermo-Story.Life (Basel, Switzerland) · 2013Review
- Structure of a periplasmic glucose-binding protein from Thermotoga maritima.Acta crystallographica. Section F, Structural biology and crystallization communications · 2012Article
- Hyperthermophilic Thermotoga species differ with respect to specific carbohydrate transporters and glycoside hydrolases.Applied and environmental microbiology · 2012Article
- Structure of the Escherichia coli phosphonate binding protein PhnD and rationally optimized phosphonate biosensors.Journal of molecular biology · 2011Article
- Ligands of thermophilic ABC transporters encoded in a newly sequenced genomic region of Thermotoga maritima MSB8 screened by differential scanning fluorimetry.Applied and environmental microbiology · 2011Article
- The L-Arabinan utilization system of Geobacillus stearothermophilus.Journal of bacteriology · 2011Article
- Engineering periplasmic ligand binding proteins as glucose nanosensors.Nano reviews · 2011Article
- Monitoring protein interactions and dynamics with solvatochromic fluorophores.Trends in biotechnology · 2010Review
- Environmentally sensitive fluorescent sensors based on synthetic peptides.Sensors (Basel, Switzerland) · 2010Review
- Comparing the performance of the optical glucose assay based on glucose binding protein with high-performance anion-exchange chromatography with pulsed electrochemical detection: efforts to design a low-cost point-of-care glucose sensor.Journal of diabetes science and technology · 2007Article
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6 authors.
Funding
Abstract
We report the design and engineering of a robust, reagentless fluorescent glucose biosensor based on the periplasmic glucose-binding protein obtained from Thermotoga maritima (tmGBP). The gene for this protein was cloned from genomic DNA and overexpressed in Escherichia coli, the identity of its cognate sugar was confirmed, ligand binding was studied, and the structure of its glucose complex was solved to 1.7 Angstrom resolution by X-ray crystallography. TmGBP is specific for glucose and exhibits high thermostability (midpoint of thermal denaturation is 119 +/- 1 degrees C and 144 +/- 2 degrees C in the absence and presence of 1 mM glucose, respectively). A series of fluorescent conjugates was constructed by coupling single, environmentally sensitive fluorophores to unique cysteines introduced by site-specific mutagenesis at positions predicted to be responsive to ligand-induced conformational changes based on the structure. These conjugates were screened to identify engineered tmGBPs that function as reagentless fluorescent glucose biosensors. The Y13C*Cy5 conjugate is bright, gives a large response to glucose over concentration ranges appropriate for in vivo monitoring of blood glucose levels (1-30 mM), and can be immobilized in an orientation-specific manner in microtiter plates to give a reversible response to glucose. The immobilized protein retains its response after long-term storage at room temperature.
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